TideCell®

The Esco TideCell® Bioreactor System is the world’s largest linearly scalable single-use bioreactor, from seed preparation to 5000 liters with closed automated cell harvesting. It is the pilot/production scale system of Esco VacciXcell’s Tide Motion Bioreactors.

One of the main advantages of Esco VacciXcell’s Tide Motion Bioreactor system is that it features the same bioprocess method from seed preparation to production scale. This is essential as certain cell lines do not grow well when seed preparation is done in 2D culture then cultured in 3D.

Esco VacciXcell's Tide Motion System is the highest yield and lowest cost packed bed bioreactor and the only packed bed bioreactor that can be placed inside a cGMP isolator, which can then be placed in Grade C-D environment, making it an ideal system for the production of wild-type viruses. This feature reduces the costs of cleanrooms by decreasing the requirements for Personnel Protective Equipment (PPE), Utilities, and Viable/Non-Viable monitoring and eliminating gowning airlocks, which are needed in larger Grade A and B cleanrooms.

No bubbling and foaming due to separate matrix vessels and mixing vessels

Low shear stress reducing cell debris

Lower downstream bioprocessing costs:

Lower cell debris in harvest medium

Much lower for secreted products, as cell remain entrapped within the carriers

No oxygen limitation due to dual oxygenation scheme

Closed system cell harvest and seed transfer

The TideCell® System is comprised of the following:

TideCell® Incubation & Control System

The TideCell® Incubation System consists of the Esco’s top of the line CO2 incubator, with additional features. It has an integrated weighing scale at the base, which allows the measurement of the amount of culture medium pumped in and out of the matrix vessel.

The TideCell® Incubation System features 2 ULPA filters. The first ULPA filter is used to ensure air quality inside the incubator, while the second ULPA filter is used to filter exhaust air, allowing the culture of BSL 3-4 virus strains. The incubation system further provides operator, environment, and sample protection with its magnetic door interlock and UV light.

Additional features of the TideCell® Incubation system include a glass window on the front door, which enables end users to observe the matrix vessel without the need to opening the incubator, reducing fluctuations in parameter values.

VacciXcell offers 2 different models of TideCell® Incubation & Control System: for 2 – 20L scale incubation & control system and 50 – 100 L scale incubation & control system. Esco’s TideCell® Incubation & Control System has user friendly design and interface and its controls are CFR 21 Part 11 compliant.

TideCell® Matrix Vessel

Single-Use Matrix VesselMultiple-Use Matrix Vessel

The matrix vessel houses the BioNOC™ II carriers, to which cells attach to and grow. VacciXcell provides a wide range of matrix vessels, depending on user’s production density requirements.

The TideCell® Mixing System holds the culture medium throughout the culture process; it is where the culture medium is continuously mixed and parameters such as pH, dissolved oxygen (DO), and temperature are monitored and regulated. This is in contrast to other bioreactor systems, where control and mixing of the culture medium takes place in the same vessel where the cells are grown.

Scada/DCS Platform as standard package runs on WonderWare Systems Platform, we are also familiar with DeltaV.

All our Tide Motion control platforms are 21 cfr part 11 compliant, following GAMP 5 guidelines and are Computer Systems Validated.

Model

Fixed-Bed Volume (L)

Culture Surface Area (m2)

Equivalent RB-850 (850 cm2)

Equivalent CS-40 (25200 cm2)

Equivalent Cytodex II (0.44 m2/8)

CelCradle

01.

1.65

20

0.65

3.75 g

TideCell-002

1~4

66

800

26

150 g

TideCell-010

5~10

165

2,000

65

375 g

TideCell-020

10~20

330

4,000

130

750 g

TideCell-100

50~100

1,650

20,000

650

3,760 g

Roller Bottles

VacciXcell’s tide motion bioreactor system is very similar to the roller bottle system in principle, as it also alternately exposes cells to aeration and nutrition. However, the tide motion system overcomes most of the disadvantages associated with the roller bottle system namely labor and space requirements and contamination risk. As costs of production become more and more expensive, existing facilities that employ the roller bottle system can thereby benefit from switching to the TideCell® System for several reasons. First, the TideCell® 100L can replace up to 20,000 850cm2 roller bottles, greatly decreasing footprint for the same productivity. Second, the TideCell® System is fully enclosed and automated, which decreases the risk of contamination and labour requirements and increases control of the production. Third, the tide motion principle employed in the TideCell® system allows simple medium exchange and better medium consumption efficiency, thereby reducing medium consumption, which is a large cost in cell culture systems. VacciXcell has successfully conducted the switch from the roller bottle system to the TideCell® system for multiple customers performing vaccine production for both human and animals.

Static Culture

Culture systems such as petri dishes, t-flasks, and cell factories employ the static principle for culture, wherein cells are continuously submerged in culture medium and exposed to aeration, without any agitation. Apart from the culture principle, the TideCell® System differs from static culture systems in terms of surface area and cell growth. Because of the fibrous macrocarriers of the TideCell® System, there is a larger surface area for cell attachment and growth in a much smaller footprint and labour requirements, as compared to static culture systems. Furthermore, this fibrous system allows cells to grow in a 3D matrix, unlike in static culture systems where cells grow in a 2D monolayer. The 3D matrix mimics cells’ in vivo environment, allowing cells to grow and develop as naturally as possible.

Microcarrier Technology

Microcarriers are microbeads, to which cells attach and grow; they are then suspended culture medium in a stirred tank bioreactor, where the medium is continuously agitated for nutrient and oxygen transfer. This agitation causes shear stress on the cells, which typically grow on the surface of the beads, causing cell death. Cell death causes 2 main problems in bioreactor systems: it causes a decrease in product yield and it increases downstream processing steps and costs due to increased cell debris in the harvest medium. The TideCell® System eliminates these problems, as cells experience negligible shear stress, while still maintaining high nutrition and oxygenation levels. In addition, scale up in a microcarrier – stirred tank bioreactor system requires much more complex parameter optimization, as compared to that of the TideCell® System, which only requires optimization of the tide motion parameters. This allows the tide motion system to have a shorter scale up bioprocessing time.

Other Packed Bed Bioreactor

In other packed bed bioreactors, cells are entrapped within a bed of carriers, while culture medium, with nutrients and dissolved oxygen, is recirculated around bed. Although these systems provide high surface area and protection from shear stress, they suffer from a critical drawback, which is oxygen limitation. Oxygen transfer in these packed reactors occur in 2 steps: air to culture medium then culture medium to cells. These two-step transfer can limit the oxygen intake of cells, thereby inhibiting cell growth. The TideCell® System overcomes oxygen limitation via its dual oxygenation scheme. First, the tide motion principle allows the direct exposure of cells to oxygen, allowing direct transfer of oxygen from the air to the cells themselves. Second, even during the nutrition phase of the tide motion, cells are still able to intake oxygen, via the dissolved oxygen in the culture medium. This dual oxygenation allows the linear scalability of the TideCell® up to 5000L.

Pilot/Production scale culture of anchorage-dependent cells

Switching from roller bottles to a closed system

Vaccine production

Allogeneic Cell Therapy

Biosimilars and Biobetters production

Cell Mass Train

Feed/Harvest Tank

Feed/Harvest system is an optional system that allows users to run the culture in perfusion/ continuous mode for long periods of time.

AutoFeeder System

AutoFeeder system has 3 major control functions: automatic control of feed and harvest volume by weight calculation, pH control, and dissolved oxygen (DO) level control.

AutoSwitch System

The AutoSwitch System is recommended if there are two feeding tanks used during fed-batch or perfusion culture and is connected in between two feeding tanks. The pinch valve inside the system will detect and automatically switch from one tank to another once the first tank is empty. An alarm light will turn on once the media in the tank is depleted; there are two individual alarm lights to indicate which feed tank which is empty. In addition, the autoswitch system includes in-line and non-invasive bubble detector.

TideCell® Cell Harvesting System

The TideCell® Cell Harvesting System (TCCHS) allows the automated and closed system harvesting of cells from the matrix vessel. TideCell® Cell Harvesting System utilizes the same conventional cell harvest concept by enzymatic treatment to detach cells from BioNOC™ II carriers.

The Tidecell® matrix vessel is transferred to the TCCHS. Harvesting protocol will be similar as user’s current harvesting protocol. Buffer is pumped in to clean out media from matrix vessel; followed by enzymes (in general, trypsin) for cell detachment, then an inhibitor (depending on the protocol). The detached cells are then collected with medium.

With this system, buffer is pumped in and out of matrix vessel 1-3 times (VacciXcell recommends to collect cells with media 5 times). This automated harvest system allows harvest cells within 1 to 3 hours, depending on harvest cycles, and shows around 95% viability after harvest. During detaching procedure, harvest equipment shakes matrix vessel at a shaking distance of 10cm and at speeds of up to 300rpm.

SCADA System

The TideCell® SCADA System enables centralized monitoring of multi-systems. The TideCell® incubation & control system, mixing system, autofeeder, and as well as autoswitch system are connected by LAN cable and monitored. With the SCADA system, users are able to access TideCell® System when it is logged in.

The SCADA system collects and records operational data such as temperature, CO2 level, pump rate; moreover, it can also record manual intervention such as who clicked fill button or who changed settings. All events and alarms that take place during the culture process are recorded and sent to the users via e-mail or other assigned communication systems.

Roller Bottles vs TideCell

Roller Bottles

TideCell

Cannot scale up only scale out which entails high space consumption and labor cost

Has contamination issues during periodic media replacement which causes very low cell yield

Fully enclosed and automatic system which greatly decreases the risk of contamination and increases production control

Microcarriers vs TideCell

Microcarriers

TideCell

Microcarriers are used for culturing adherent cells in suspension. Agitation applied on the microcarrier-bound-cells causes great shear stress which leads to cell death or apoptosis

Due to tide motion principle TideCell will have negligible shear stress while maintaining high nutrition and oxygenation levels which results to high and more concentrated cell yield

Using microcarriers increases downstream processing steps and costs due to increased cell debris and difficulty in the separation of the cells from the microcarriers

Minimal downstream processing since cells are attached to BioNocII macrocarriers while cell debris and other impurities can easily be removed with the media

Complex parameter optimization upon scale up

True Linear Scalability

Other Packed Bed Bioreactors vs TideCell

Other Packed Bed Bioreactors

TideCell

Oxygenation is reliant on dissolved oxygen in media, making O2 intake of cells uneven and limited, inhibiting cell growth

TideCell’s dual oxygenation system provides every cell within the matrix ample supply of oxygen regardless of the size or scale of the vessel

Limited oxygenation inhibits scale up

Macroporous carriers used in other packed bed bioreactors are made of only 50% PET and 50% PP

BioNocII which is made of 100% PET ensures that cells adhere completely to the macrocarriers and as a result high product and cell yield can be achieved.

The following table shows cGMP compliant customers using TideCell in their projects. Company identities and some other information are hidden due to Non-disclosure agreement (NDA) between the company and VacciXcell

The TideCell® Incubation System consists of the Esco’s top of the line CO2 incubator, with additional features. It has an integrated weighing scale at the base, which allows the measurement of the amount of culture medium pumped in and out of the matrix vessel.

The TideCell® Incubation System features 2 ULPA filters. The first ULPA filter is used to ensure air quality inside the incubator, while the second ULPA filter is used to filter exhaust air, allowing the culture of BSL 3-4 virus strains. The incubation system further provides operator, environment, and sample protection with its magnetic door interlock and UV light.

Additional features of the TideCell® Incubation system include a glass window on the front door, which enables end users to observe the matrix vessel without the need to opening the incubator, reducing fluctuations in parameter values.

VacciXcell offers 2 different models of TideCell® Incubation & Control System: for 2 – 20L scale incubation & control system and 50 – 100 L scale incubation & control system. Esco’s TideCell® Incubation & Control System has user friendly design and interface and its controls are CFR 21 Part 11 compliant.

TideCell® Matrix Vessel

Single-Use Matrix VesselMultiple-Use Matrix Vessel

The matrix vessel houses the BioNOC™ II carriers, to which cells attach to and grow. VacciXcell provides a wide range of matrix vessels, depending on user’s production density requirements.

The TideCell® Mixing System holds the culture medium throughout the culture process; it is where the culture medium is continuously mixed and parameters such as pH, dissolved oxygen (DO), and temperature are monitored and regulated. This is in contrast to other bioreactor systems, where control and mixing of the culture medium takes place in the same vessel where the cells are grown.

VacciXcell’s tide motion bioreactor system is very similar to the roller bottle system in principle, as it also alternately exposes cells to aeration and nutrition. However, the tide motion system overcomes most of the disadvantages associated with the roller bottle system namely labor and space requirements and contamination risk. As costs of production become more and more expensive, existing facilities that employ the roller bottle system can thereby benefit from switching to the TideCell® System for several reasons. First, the TideCell® 100L can replace up to 20,000 850cm2 roller bottles, greatly decreasing footprint for the same productivity. Second, the TideCell® System is fully enclosed and automated, which decreases the risk of contamination and labour requirements and increases control of the production. Third, the tide motion principle employed in the TideCell® system allows simple medium exchange and better medium consumption efficiency, thereby reducing medium consumption, which is a large cost in cell culture systems. VacciXcell has successfully conducted the switch from the roller bottle system to the TideCell® system for multiple customers performing vaccine production for both human and animals.

Static Culture

Culture systems such as petri dishes, t-flasks, and cell factories employ the static principle for culture, wherein cells are continuously submerged in culture medium and exposed to aeration, without any agitation. Apart from the culture principle, the TideCell® System differs from static culture systems in terms of surface area and cell growth. Because of the fibrous macrocarriers of the TideCell® System, there is a larger surface area for cell attachment and growth in a much smaller footprint and labour requirements, as compared to static culture systems. Furthermore, this fibrous system allows cells to grow in a 3D matrix, unlike in static culture systems where cells grow in a 2D monolayer. The 3D matrix mimics cells’ in vivo environment, allowing cells to grow and develop as naturally as possible.

Microcarrier Technology

Microcarriers are microbeads, to which cells attach and grow; they are then suspended culture medium in a stirred tank bioreactor, where the medium is continuously agitated for nutrient and oxygen transfer. This agitation causes shear stress on the cells, which typically grow on the surface of the beads, causing cell death. Cell death causes 2 main problems in bioreactor systems: it causes a decrease in product yield and it increases downstream processing steps and costs due to increased cell debris in the harvest medium. The TideCell® System eliminates these problems, as cells experience negligible shear stress, while still maintaining high nutrition and oxygenation levels. In addition, scale up in a microcarrier – stirred tank bioreactor system requires much more complex parameter optimization, as compared to that of the TideCell® System, which only requires optimization of the tide motion parameters. This allows the tide motion system to have a shorter scale up bioprocessing time.

Other Packed Bed Bioreactor

In other packed bed bioreactors, cells are entrapped within a bed of carriers, while culture medium, with nutrients and dissolved oxygen, is recirculated around bed. Although these systems provide high surface area and protection from shear stress, they suffer from a critical drawback, which is oxygen limitation. Oxygen transfer in these packed reactors occur in 2 steps: air to culture medium then culture medium to cells. These two-step transfer can limit the oxygen intake of cells, thereby inhibiting cell growth. The TideCell® System overcomes oxygen limitation via its dual oxygenation scheme. First, the tide motion principle allows the direct exposure of cells to oxygen, allowing direct transfer of oxygen from the air to the cells themselves. Second, even during the nutrition phase of the tide motion, cells are still able to intake oxygen, via the dissolved oxygen in the culture medium. This dual oxygenation allows the linear scalability of the TideCell® up to 5000L.

Feed/Harvest system is an optional system that allows users to run the culture in perfusion/ continuous mode for long periods of time.

AutoFeeder System

AutoFeeder system has 3 major control functions: automatic control of feed and harvest volume by weight calculation, pH control, and dissolved oxygen (DO) level control.

AutoSwitch System

The AutoSwitch System is recommended if there are two feeding tanks used during fed-batch or perfusion culture and is connected in between two feeding tanks. The pinch valve inside the system will detect and automatically switch from one tank to another once the first tank is empty. An alarm light will turn on once the media in the tank is depleted; there are two individual alarm lights to indicate which feed tank which is empty. In addition, the autoswitch system includes in-line and non-invasive bubble detector.

TideCell® Cell Harvesting System

The TideCell® Cell Harvesting System (TCCHS) allows the automated and closed system harvesting of cells from the matrix vessel. TideCell® Cell Harvesting System utilizes the same conventional cell harvest concept by enzymatic treatment to detach cells from BioNOC™ II carriers.

The Tidecell® matrix vessel is transferred to the TCCHS. Harvesting protocol will be similar as user’s current harvesting protocol. Buffer is pumped in to clean out media from matrix vessel; followed by enzymes (in general, trypsin) for cell detachment, then an inhibitor (depending on the protocol). The detached cells are then collected with medium.

With this system, buffer is pumped in and out of matrix vessel 1-3 times (VacciXcell recommends to collect cells with media 5 times). This automated harvest system allows harvest cells within 1 to 3 hours, depending on harvest cycles, and shows around 95% viability after harvest. During detaching procedure, harvest equipment shakes matrix vessel at a shaking distance of 10cm and at speeds of up to 300rpm.

SCADA System

The TideCell® SCADA System enables centralized monitoring of multi-systems. The TideCell® incubation & control system, mixing system, autofeeder, and as well as autoswitch system are connected by LAN cable and monitored. With the SCADA system, users are able to access TideCell® System when it is logged in.

The SCADA system collects and records operational data such as temperature, CO2 level, pump rate; moreover, it can also record manual intervention such as who clicked fill button or who changed settings. All events and alarms that take place during the culture process are recorded and sent to the users via e-mail or other assigned communication systems.

The following table shows cGMP compliant customers using TideCell in their projects. Company identities and some other information are hidden due to Non-disclosure agreement (NDA) between the company and VacciXcell